Search results for: natural killer (NK) cells
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 8498

Search results for: natural killer (NK) cells

7658 Cryogenic Separation of CO2 from Molten Carbonate Fuel Cell Anode Outlet—Experimental Guidelines

Authors: Jarosław Milewski, Rafał Bernat

Abstract:

This paper presents an analysis of using cryogenic separation unit for recovering fuel from anode off gas of molten carbonate fuel cells (MCFCs) in order to upgrade the efficiently of the unit. In the proposed solution, the CSU is used for condensing water and carbon dioxide from anode off gas, and re-cycling the rest of the stream to the anode, saving certain amount of fuel (at least 30%). The resulting system efficiency is increased considerably. CSU, virtually consumes power, thus this solution has energy penalty as well, on the other hand, MCFC generates large amount of heat at elevated temperature, thus part of the CSU can be based on absorption chiller. In all cases, a high amount of fuel is obtained after condensation of water and carbon dioxide and re-cycled to the anode inlet. Based on mathematical modeling done previously, the concept and guidelines for forthcoming experimental investigations are presented in this paper. During planned experiments, an existing single cell laboratory stand will be equipped with re-cycle device (a fan, a peristaltic pump, etc.). Parallel, a mixture of anode off gas will be cooled down for determining the proper temperature for the separation of water and carbon dioxide.

Keywords: cryogenic separation, experiments, fuel cells, molten carbonate fuel cells

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7657 Furnishing The Envelope; 3D Printed Construction Unit as Furniture

Authors: Maryam Kalkatechi

Abstract:

The paper presents the construction unit that was proposed as a result of researching and finding solutions for challenges of the traditional masonry unit. The concept of ‘unit as arrangements of cells’ was investigated in four categories of structure, handling and assembly, thermal characteristics and weather ability which resulted in construction unit as an independent system which shapes a part of the envelope. Comparing to the traditional wall systems in which the system is in layers, the part system is a monolithic piece by itself. Even though the overall wythe-10 inches- is less than the combined layers-14 inches- in a traditional wall system, it is still seen as a spatial component. The component as a furnishing of envelope is discussed from material application point of view. The algorithm definition of the arrangement cells crafts the relationship between cells and functionality with material. This craft is realized as the envelope furnishing. Three alternative materials in relation to furnishing the envelope are discussed for printing the construction unit; transparent plastic, opaque plastic and glass. The qualities vary in the four categories, however this paper focuses on the visual qualities of materials applied. In a diagram the qualities of the materials are compared in relation to each other.

Keywords: furnishing envelope, 3D printed construction unit, opaque plastic, transparent plastic, glass

Procedia PDF Downloads 165
7656 In vitro Culture of Stem Node Segments of Maerua crassifolia

Authors: Abobaker Abrahem M. Saad, Asma Abudasalam

Abstract:

The stem node segments were cultured on Murashige and Skoog (MS) medium. In the case of using MS+ Zeatin (1 mg/l), small shoot buds were formed directly in 70% of explants after 15 days, their length range between 0.1 to 0.3 cm after two weeks and reached 0.3 cm in length and three shoots in numbers after 4 weeks. When those small shoots were sub cultured on the same medium, they increased in length, number and reached 0.4 cm with 4 shoots, 0.4 cm with 5 shoots after six, eight and ten weeks respectively. In the case of using MS free hormones, MS+IAA (0.2mg/l) +BA (0.5mg/l), MS + kin(0.5mg/l), MS + kin (3mg/l) and MS +NAA (3mg/l) +BA (1mg/l), no sign of responses were noticed and only change in color in some cases. Different types of parenchyma cells and many layers of thick wall sclerenchyma cells were observed on MS+BA (1mg/l).

Keywords: Maerua, stem node, shoots, buds, In vitro

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7655 Undifferentiated Embryonal Sarcoma of Liver: A Rare Case Report

Authors: Thieu-Thi Tra My

Abstract:

Undifferentiated embryonal sarcoma of the liver (UESL), a rare malignant mesenchymal tumor, is commonly seen in children. The symptoms and imaging were not specific, so it could be mimicked with other tumors or liver abscesses. The tumor often appears as a large heterogeneous echoic solid mass with small cystic areas while showing a cyst-like appearance on CT and MRI. The histopathological manifestation of the UESL consisted of stellate-shaped and spindle cells scattered on a myxoid background with high mitotic count. Cells with multiple or bizarre nuclear were also observed. Here, we aimed to describe a 9-year-old male diagnosed with UESL focused on imaging and histopathological characteristics.

Keywords: undifferentiated embryonal sarcoma of liver, UESL, liver sarcoma, liver tumor, children

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7654 Predicting of Hydrate Deposition in Loading and Offloading Flowlines of Marine CNG Systems

Authors: Esam I. Jassim

Abstract:

The main aim of this paper is to demonstrate the prediction of the model capability of predicting the nucleation process, the growth rate, and the deposition potential of second phase particles in gas flowlines. The primary objective of the research is to predict the risk hazards involved in the marine transportation of compressed natural gas. However, the proposed model can be equally used for other applications including production and transportation of natural gas in any high-pressure flow-line. The proposed model employs the following three main components to approach the problem: computational fluid dynamics (CFD) technique is used to configure the flow field; the nucleation model is developed and incorporated in the simulation to predict the incipient hydrate particles size and growth rate; and the deposition of the gas/particle flow is proposed using the concept of the particle deposition velocity. These components are integrated in a comprehended model to locate the hydrate deposition in natural gas flowlines. The present research is prepared to foresee the deposition location of solid particles that could occur in a real application in Compressed Natural Gas loading and offloading. A pipeline with 120 m length and different sizes carried a natural gas is taken in the study. The location of particle deposition formed as a result of restriction is determined based on the procedure mentioned earlier and the effect of water content and downstream pressure is studied. The critical flow speed that prevents such particle to accumulate in the certain pipe length is also addressed.

Keywords: hydrate deposition, compressed natural gas, marine transportation, oceanography

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7653 Researches on Attractive Flowered Natural Woody Plants of Bursa Flora in Terms of Landscape Design

Authors: Elvan Ender, Murat Zencirkıran

Abstract:

One of the most important criteria that increase the success of design in landscape architecture is the visual effect. The characteristics that affect visual appearance in plant design vary depending on the phenological periods of the plants. In plants, although different effects are observed in different periods of the year, this effect is felt most prominently in flowering periods. For this reason, knowing the flowering time, duration and flower characteristics should be considered as a factor increasing the success of plant design. In this study, flower characteristics of natural woody plants with attractive flowers have been examined. Because of the variability of these characteristics of plants in the region, consideration of these criteria in the planting design processes in the region may increase the success of the design. At the same time, when species selection is made considering the obtained data, visuality and sustainability of natural species can be possible in Bursa city with planting design.

Keywords: Bursa, flower characteristics, natural plants, planting design

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7652 The Molecular Rationale for Steroid Based Therapy of Leukemia: Diagnostic and Therapeutic Implications

Authors: Eitan Yefenof

Abstract:

Glucocorticoid (GC) hormones, e.g. Dexamethasone and Prednisone, are widely used in the therapy of leukemia and lymphoma owing to their apoptogenic effect on lymphoid cells. However, the emergence of GC resistant cells during therapy is a major cause for treatment failure, urging the need for novel strategies that maintain leukemia sensitivity to the pro-apoptotic activity of GCs. GCs act by binding to the GC receptor (GR), which, in its inactive state, is sequestered in the cytosol by a multi-subunit complex of heat shock proteins. Upon ligand binding, the complex dissociates, allowing GR activation and translocation to the nucleus, where it regulates transcription of multiple genes. We demonstrated that in addition to gene expression, GR also regulates microRNA (miR) expression. Deep-sequencing analysis revealed 14 miRs that are regulated in GC-sensitive but resistant leukemias upon treatment with GC. GC up-regulates miR-103, miR-15~16 and miR-30e/d, while down-regulates miR-17, mir-18a, miR-19a, miR-19b, miR-20a and miR-92a (members of the miR-17∼92a multi-cistron). Upon transfection, miR-103 confers GC apoptotic sensitivity to otherwise GC-resistant cell. Furthermore, knocking down miR-103 expression reduces the GC apoptotic response of sensitive cells. miR-103 abrogates c-Myc expression, an oncogenic transcription factor which is deregulated in many cancers. In addition, miR-103 up-regulates Bim, a pro-apoptotic protein crucial for GC-induced death. Activated glycogen synthase kinase 3 (GSK3) is also crucial for GC-induced apoptosis. GSK3 is active in GC-sensitive but not in GC-resistant cells. We found that GSK3 associates with the GR multi-subunit complex. Upon GC exposure, it dissociates from the GR and interacts with Bim to enable activation of the mitochondrial apoptosis pathway. miR-103 mediated c-Myc ablation is followed by down-regulation of the multi-cistron miR-17~92a, in particular miR-18a and miR-20a. miR-18a targets GR for degradation whereas miR-20a targets Bim degradation. Hence, miR-103 acts, in concert with Bim and GR, as a "tumor suppressor" that leads to reduced proliferation, cell-cycle arrest and cell death. We suggest that miR-103 can provide a diagnostic tool that predicts the sensitivity of leukemia to GC based therapy. Furthermore, exosomal delivery of miR-103 or up-regulation of the endogenous miR-103 could confer apoptotic sensitivity to resistant cells at the outset, thus becoming a useful therapeutic tool combined with GCs.

Keywords: apoptosis, leukemia, micro-RNA, steroids

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7651 Effect of He-Ne Laser Therapy on the Testis and Serum Testosterone Level in Adult Rats

Authors: Nadeem H. Meikha , Nazad H. Qader, Basheer M. Hasafa

Abstract:

The trial was conducted to examine the effect of He-Ne laser therapy on the testis and serum testosterone level in adult rats. Thirty five albino Western adult male rats aged 3-4 months and weighing approximately 250-300 g were used and divided into three treatments. Testicular tissue of rats in the first and second treatments were exposed once daily for three successively days to a dose of irradiation 1.02 j/cm2 (40 second), and to 2.03 j/cm2 (80 second) respectively, while the third group left without any treatments (control). The results showed that the process of irradiation adversely affected on the level of serum testosterone concentration of the irradiated rats in the first and second treatment comparing to the normal level in the control group. While the histological examination showed that decrease in number of germ cells with 40 second of irradiation at day three, with 80 second of irradiation the decreased started at day two and three. The spermatids number decreased in rate low, medium, high respectively for three days of 40 second of irradiation, while the spermatids number were adversely affected by dropping in a rate of medium, large and very large for three days of 80 second of irradiation, respectively. In conclusion our study revealed that any reduction in sertoli cells causes adverse affect on both spermatids and germinal cells which increase with the increasing of duration and repetition of irradiation.

Keywords: He-Ne laser, rats, testosterone, spermatids

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7650 Growth and Differentiation of Mesenchymal Stem Cells on Titanium Alloy Ti6Al4V and Novel Beta Titanium Alloy Ti36Nb6Ta

Authors: Eva Filová, Jana Daňková, Věra Sovková, Matej Daniel

Abstract:

Titanium alloys are biocompatible metals that are widely used in clinical practice as load bearing implants. The chemical modification may influence cell adhesion, proliferation, and differentiation as well as stiffness of the material. The aim of the study was to evaluate the adhesion, growth and differentiation of pig mesenchymal stem cells on the novel beta titanium alloy Ti36Nb6Ta compared to standard medical titanium alloy Ti6Al4V. Discs of Ti36Nb6Ta and Ti6Al4V alloy were sterilized by ethanol, put in 48-well plates, and seeded by pig mesenchymal stem cells at the density of 60×103/cm2 and cultured in Minimum essential medium (Sigma) supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cell viability was evaluated using MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay;Promega), cell proliferation using Quant-iT™ ds DNA Assay Kit (Life Technologies). Cells were stained immunohistochemically using monoclonal antibody beta-actin, and secondary antibody conjugated with AlexaFluor®488 and subsequently the spread area of cells was measured. Cell differentiation was evaluated by alkaline phosphatase assay using p-nitrophenyl phosphate (pNPP) as a substrate; the reaction was stopped by NaOH, and the absorbance was measured at 405 nm. Osteocalcin, specific bone marker was stained immunohistochemically and subsequently visualized using confocal microscopy; the fluorescence intensity was analyzed and quantified. Moreover, gene expression of osteogenic markers osteocalcin and type I collagen was evaluated by real-time reverse transcription-PCR (qRT-PCR). For statistical evaluation, One-way ANOVA followed by Student-Newman-Keuls Method was used. For qRT-PCR, the nonparametric Kruskal-Wallis Test and Dunn's Multiple Comparison Test were used. The absorbance in MTS assay was significantly higher on titanium alloy Ti6Al4V compared to beta titanium alloy Ti36Nb6Ta on days 7 and 14. Mesenchymal stem cells were well spread on both alloys, but no difference in spread area was found. No differences in alkaline phosphatase assay, fluorescence intensity of osteocalcin as well as the expression of type I collagen, and osteocalcin genes were observed. Higher expression of type I collagen compared to osteocalcin was observed for cells on both alloys. Both beta titanium alloy Ti36Nb6Ta and titanium alloy Ti6Al4V Ti36Nb6Ta supported mesenchymal stem cellsˈ adhesion, proliferation and osteogenic differentiation. Novel beta titanium alloys Ti36Nb6Ta is a promising material for bone implantation. The project was supported by the Czech Science Foundation: grant No. 16-14758S, the Grant Agency of the Charles University, grant No. 1246314 and by the Ministry of Education, Youth and Sports NPU I: LO1309.

Keywords: beta titanium, cell growth, mesenchymal stem cells, titanium alloy, implant

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7649 The Activity of Polish Propolis and Cannabidiol Oil Extracts on Glioblastoma Cell Lines

Authors: Sylwia K. Naliwajko, Renata Markiewicz-Zukowska, Justyna Moskwa, Krystyna Gromkowska-Kepka, Konrad Mielcarek, Patryk Nowakowski, Katarzyna Socha, Anna Puscion-Jakubik, Maria H. Borawska

Abstract:

Glioblastoma (grade IV WHO) is a rapidly progressive brain tumor with very high morbidity and mortality. The vast malignant gliomas are not curable despite the therapy (surgical, radiotherapy, chemotherapy) and patients seek alternative or complementary treatments. Patients often use cannabidiol (CBD) oil as an alternative therapy of glioblastoma. CBD is one of the cannabinoids, an active component of Cannabis sativa. THC (Δ9-tetrahydrocannabinol) can be addictive, and in many countries CBD oil without THC ( < 0,2%) is available. Propolis produced by bees from the resin collected from trees has antiglioma properties in vitro and can be used as a supplement in complementary therapy of gliomas. The aim of this study was to examine the influence of extract from CBD oil in combination with propolis extract on two glioblastoma cell lines. The MTT (Thiazolyl Blue Tetrazolium Bromide) test was used to determine the influence of CBD oil extract and polish propolis extract (PPE) on the viability of glioblastoma cell lines – U87MG and LN18. The cells were incubated (24, 48 and 72 h) with CBD oil extract and PPE. CBD extract was used in concentration 1, 1.5 and 3 µM and PPE in 30 µg/mL. The data were presented compared to the control. The statistical analysis was performed using Statistica v. 13.0 software. CBD oil extract in concentrations 1, 1.5 and 3 µM did not inhibit the viability of U87MG and LN18 cells (viability more than 90% cells compared to the control). There was no dose-response viability, and IC50 value was not recognized. PPE in the concentration of 30 µg/mL time-dependently inhibited the viability of U87MG and LN18 cell line (after 48 h the viability as a percent of the control was 59,7±6% and 57,8±7%, respectively). In a combination of CBD with PPE, the viability of the treated cells was similar to PPE used alone (58,2±7% and 56,5±9%, respectively). CBD oil extract did not show anti-glioma activity and in combination with PPE did not change the activity of PPE.

Keywords: anticancer, cannabidiol, cell line, glioblastoma

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7648 Comparative Study of Calcium Content on in vitro Biological and Antibacterial Properties of Silicon-Based Bioglass

Authors: Morteza Elsa, Amirhossein Moghanian

Abstract:

The major aim of this study was to evaluate the effect of CaO content on in vitro hydroxyapatite formation, MC3T3 cells cytotoxicity and proliferation as well as antibacterial efficiency of sol-gel derived SiO2–CaO–P2O5 ternary system. For this purpose, first two grades of bioactive glass (BG); BG-58s (mol%: 60%SiO2–36%CaO–4%P2O5) and BG-68s (mol%: 70%SiO2–26%CaO–4%P2O5)) were synthesized by sol-gel method. Second, the effect of CaO content in their composition on in vitro bioactivity was investigated by soaking the BG-58s and BG-68s powders in simulated body fluid (SBF) for time periods up to 14 days and followed by characterization inductively coupled plasma atomic emission spectrometry (ICP-AES), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM) techniques. Additionally, live/dead staining, 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and alkaline phosphatase (ALP) activity assays were conducted respectively, as qualitatively and quantitatively assess for cell viability, proliferation and differentiations of MC3T3 cells in presence of 58s and 68s BGs. Results showed that BG-58s with higher CaO content showed higher in vitro bioactivity with respect to BG-68s. Moreover, the dissolution rate was inversely proportional to oxygen density of the BG. Live/dead assay revealed that both 58s and 68s increased the mean number live cells which were in good accordance with MTT assay. Furthermore, BG-58s showed more potential antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) bacteria. Taken together, BG-58s with enhanced MC3T3 cells proliferation and ALP activity, acceptable bioactivity and significant high antibacterial effect against MRSA bacteria is suggested as a suitable candidate in order to further functionalizing for delivery of therapeutic ions and growth factors in bone tissue engineering.

Keywords: antibacterial, bioactive glass, hydroxyapatite, proliferation, sol-gel processes

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7647 Cytotoxicological Evaluation of a Folate Receptor Targeting Drug Delivery System Based on Cyclodextrins

Authors: Caroline Mendes, Mary McNamara, Orla Howe

Abstract:

For chemotherapy, a drug delivery system should be able to specifically target cancer cells and deliver the therapeutic dose without affecting normal cells. Folate receptors (FR) can be considered key targets since they are commonly over-expressed in cancer cells and they are the molecular marker used in this study. Here, cyclodextrin (CD) has being studied as a vehicle for delivering the chemotherapeutic drug, methotrexate (MTX). CDs have the ability to form inclusion complexes, in which molecules of suitable dimensions are included within the CD cavity. In this study, β-CD has been modified using folic acid so as to specifically target the FR molecular marker. Thus, the system studied here for drug delivery consists of β-CD, folic acid and MTX (CDEnFA:MTX). Cellular uptake of folic acid is mediated with high affinity by folate receptors while the cellular uptake of antifolates, such as MTX, is mediated with high affinity by the reduced folate carriers (RFCs). This study addresses the gene (mRNA) and protein expression levels of FRs and RFCs in the cancer cell lines CaCo-2, SKOV-3, HeLa, MCF-7, A549 and the normal cell line BEAS-2B, quantified by real-time polymerase chain reaction (real-time PCR) and flow cytometry, respectively. From that, four cell lines with different levels of FRs, were chosen for cytotoxicity assays of MTX and CDEnFA:MTX using the MTT assay. Real-time PCR and flow cytometry data demonstrated that all cell lines ubiquitously express moderate levels of RFC. These experiments have also shown that levels of FR protein in CaCo-2 cells are high, while levels in SKOV-3, HeLa and MCF-7 cells are moderate. A549 and BEAS-2B cells express low levels of FR protein. FRs are highly expressed in all the cancer cell lines analysed when compared to the normal cell line BEAS-2B. The cell lines CaCo-2, MCF-7, A549 and BEAS-2B were used in the cell viability assays. 48 hours treatment with the free drug and the complex resulted in IC50 values of 93.9 µM ± 9.2 and 56.0 µM ± 4.0 for CaCo-2 for free MTX and CDEnFA:MTX respectively, 118.2 µM ± 10.8 and 97.8 µM ± 12.3 for MCF-7, 36.4 µM ± 6.9 and 75.0 µM ± 8.5 for A549 and 132.6 µM ± 12.1 and 288.1 µM ± 16.3 for BEAS-2B. These results demonstrate that MTX is more toxic towards cell lines expressing low levels of FR, such as the BEAS-2B. More importantly, these results demonstrate that the inclusion complex CDEnFA:MTX showed greater cytotoxicity than the free drug towards the high FR expressing CaCo-2 cells, indicating that it has potential to target this receptor, enhancing the specificity and the efficiency of the drug.

Keywords: cyclodextrins, cancer treatment, drug delivery, folate receptors, reduced folate carriers

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7646 Targeting the EphA2 Receptor Tyrosine Kinases in Melanoma Cancer, both in Humans and Dogs

Authors: Shabnam Abdi, Behzad Toosi

Abstract:

Background: Melanoma is the most lethal type of malignant skin cancer in humans and dogs since it spreads rapidly throughout the body. Despite significant advances in treatment, cancer at an advanced stage has a poor prognosis. Hence, more effective treatments are needed to enhance outcomes with fewer side effects. Erythropoietin-producing hepatocellular receptors are the largest family of receptor tyrosine kinases and are divided into two subfamilies, EphA and EphB, both of which play a significant role in disease, especially cancer. Due to their association with proliferation and invasion in many aggressive types of cancer, Eph receptor tyrosine kinases (Eph RTKs) are promising cancer therapy molecules. Because these receptors have not been studied in canine melanoma, we investigated how EphA2 influences survival and tumorigenicity of melanoma cells. Methods: Expression of EphA2 protein in canine melanoma cell lines and human melanoma cell line was evaluated by Western blot. Melanoma cells were transduced with lentiviral particles encoding Eph-targeting shRNAs or non-silencing shRNAs (control) for silencing the expression of EphA2 receptor, and silencing was confirmed by Western blotting and immunofluorescence. The effect of siRNA treatment on cellular proliferation, colony formation, tumorsphere assay, invasion was analyzed by Resazurin assay Matrigel invasion assay, respectively. Results: Expression of EphA2 was detected in canine and human melanoma cell lines. Moreover, stably silencing EphA2 by specific shRNAs significantly and consistently decreased the expression of EphA2 protein in both human and canine melanoma cells. Proliferation, colony formation, tumorsphere and invasion of melanoma cells were significantly decreased in EphA2 siRNA-treated cells compared to control. Conclusion: Our data provide the first functional evidence that the EphA2 receptor plays a critical role in the malignant cellular behavior of melanoma in both human and dogs.

Keywords: ephA2, targeting, melanoma, human, canine

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7645 Effect of Mineral Additives on Improving the Geotechnical Properties of Soils in Chlef

Authors: Messaoudi Mohammed Amin

Abstract:

The reduction of available land resources and the increased cout associated with the use of hight quality materials have led to the need for local soils to be used in geotecgnical construction however, poor engineering properties of these soils pose difficulties for constructions project and need to be stabilized to improve their properties in oyher works unsuitable soils with low bearing capacity, high plasticity coupled with high insatbility are frequently encountered hense, there is a need to improve the physical and mechanical charateristics of these soils to make theme more suitable for construction this can be done by using different mechanical and chemical methods clayey soil stabilization has been practiced for quite sometime bu mixing additives, such us cement, lime and fly ash to the soil to increase its strength. The aim of this project is to study the effect of using lime, natural pozzolana or combination of both on the geotecgnical cherateristics of clayey soil. Test specimen were subjected to atterberg limits test, compaction test, box shear test and uncomfined compression test Lime or natural pozzolana was added to clayey soil at rangs of 0-8% and 0-20% respectively. In addition combinations of lime –natural pozzolana were added to clayey soil at the same ranges specimen were cured for 1-7, and 28 days after which they were tested for uncofined compression tests. Based on the experimental results, it was concluded that an important decrease of plasticity index was observed for thr samples stabilized with the combinition lime-natural pozzolana in addition, the use of the combination lime-natural pozzolana modifies the clayey soil classification according to casagrand plasiticity chart. Moreover, based on the favourable results of shear and compression strength obtained, it can be concluded that clayey soil can be successfuly stabilized by combined action of lime and natural pozzolana also this combination showed an appreciable improvement of the shear parameters. Finally, since natural pozzolana is much cheaper than lime ,the addition of natural pozzolana in lime soil mix may particulary become attractive and can result in cost reduction of construction.

Keywords: clay, soil stabilization, natural pozzolana, atterberg limits, compaction, compressive strength shear strength, curing

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7644 Determination of Natural Gamma Radioactivity in Sand along the Black Sea Coastal Region of Giresun, North Turkey

Authors: A. Karadeniz, Belgin Kucukomeroglu

Abstract:

In this study natural gamma radioactivity levels are determined on sands along the coastal regions of Giresun/Turkey. The coast of Giresun about 290 km long in investigated to collect 101 sand samples. Natural and artificial radioactivity concentrations of sand samples were measured by using HPGe gamma spectrometry. The average activity concentrations of 238U, 232Th, 40K and 137Cs on sand samples of Giresun were found to be 10.83±2.92 Bq/kg, 21.28±3.22 Bq/kg, 6.42±1.06 Bq/kg, 230.94±10.67 Bq/kg respectively. The average activity concentrations for these radionuclides were compared with the reported data of other parts of Turkey and other countries. The average absorbed dose rate for Giresun was calculated to be 38.68 nGy/h respectively. This value is significantly lower than the World averaged value of 60 nGy/h. The external annual effective dose rate concentration in Giresun was found to be 0.047 mSv/y respectively. This result is much lower than the recommeded limit of 5 mSv/y. The external hazard dose rate for Giresun weas calculated to be 0.21 respectively. This result is much lower than the recommended limit of 1.0.

Keywords: concentration, radioactivity, Giresun, natural gamma radioactivity

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7643 Preparation of Allyl BODIPY for the Click Reaction with Thioglycolic Acid

Authors: Chrislaura Carmo, Luca Deiana, Mafalda Laranjo, Abilio Sobral, Armando Cordova

Abstract:

Photodynamic therapy (PDT) is currently used for the treatment of malignancies and premalignant tumors. It is based on the capture of a photosensitizing molecule (PS) which, when excited by light at a certain wavelength, reacts with oxygen and generates oxidizing species (radicals, singlet oxygen, triplet species) in target tissues, leading to cell death. BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indaceno) derivatives are emerging as important candidates for photosensitizer in photodynamic therapy of cancer cells due to their high triplet quantum yield. Today these dyes are relevant molecules in photovoltaic materials and fluorescent sensors. In this study, it will be demonstrated the possibility that BODIPY can be covalently linked to thioglycolic acid through the click reaction. Thiol−ene click chemistry has become a powerful synthesis method in materials science and surface modification. The design of biobased allyl-terminated precursors with high renewable carbon content for the construction of the thiol-ene polymer networks is essential for sustainable development and green chemistry. The work aims to synthesize the BODIPY (10-(4-(allyloxy) phenyl)-2,8-diethyl-5,5-difluoro-1,3,7,9-tetramethyl-5H-dipyrrolo[1,2-c:2',1'-f] [1,3,2] diazaborinin-4-ium-5-uide) and to click reaction with Thioglycolic acid. BODIPY was synthesized by the condensation reaction between aldehyde and pyrrole in dichloromethane, followed by in situ complexation with BF3·OEt2 in the presence of the base. Then it was functionalized with allyl bromide to achieve the double bond and thus be able to carry out the click reaction. The thiol−ene click was performed using DMPA (2,2-Dimethoxy-2-phenylacetophenone) as a photo-initiator in the presence of UV light (320–500 nm) in DMF at room temperature for 24 hours. Compounds were characterized by standard analytical techniques, including UV-Vis Spectroscopy, 1H, 13C, 19F NMR and mass spectroscopy. The results of this study will be important to link BODIPY to polymers through the thiol group offering a diversity of applications and functionalization. This new molecule can be tested as third-generation photosensitizers, in which the dye is targeted by antibodies or nanocarriers by cells, mainly in cancer cells, PDT and Photodynamic Antimicrobial Chemotherapy (PACT). According to our studies, it was possible to visualize a click reaction between allyl BODIPY and thioglycolic acid. Our team will also test the reaction with other thiol groups for comparison. Further, we will do the click reaction of BODIPY with a natural polymer linked with a thiol group. The results of the above compounds will be tested in PDT assays on various lung cancer cell lines.

Keywords: bodipy, click reaction, thioglycolic acid, allyl, thiol-ene click

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7642 Concrete Performance Evaluation of Coarse Aggregate Replacement by Civil Construction Waste

Authors: Juliane P. De Oliveira, Carlos H. Dos Santos, Marcia Shoji, Maria E. C. Ferreira, Natalia U. Yamaguchi

Abstract:

The construction sector is considered a major generator of environmental impacts due to the high consumption of natural resources and waste generation. Thus, this article aims to evaluate the performance of a concrete produced by the partial and total replacement of natural coarse aggregate by recycled coarse aggregate, derived from the concrete residue of buildings and demolitions. The study was made by comparing the compressive strength and absorption of three different concrete traces, keeping the water/cement factor of 0.60 and changing only the proportions of recycled coarse aggregate between 0%, 50% and 100%. Traces 50% and 100% obtained good results by comparing the actual specific mass, because the material used is lighter to the natural coarse aggregate. It was concluded that the concrete produced with recycled aggregates, even with inferior results, can be used where it is not needed a structural function, giving an adequate destination to the construction and demolition waste and consequently reducing the extraction and consumption of natural resources.

Keywords: green concrete, recycled aggregate, recycling, sustainable development

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7641 Fuel Cells and Offshore Wind Turbines Technology for Eco-Friendly Ports with a Case Study

Authors: Ibrahim Sadek Sedik Ibrahim, Mohamed M. Elgohary

Abstract:

Sea ports are considered one of the factors affecting the progress of economic globalization and the international trade; consequently, they are considered one of the sources involved in the deterioration of the maritime environment due to the excessive amount of exhaust gases emitted from their activities. The majority of sea ports depend on the national electric grid as a source of power for the domestic and ships’ electric demands. This paper discusses the possibility of shifting ports from relying on the national grid electricity to green power-based ports. Offshore wind turbines and hydrogenic PEM fuel cell units appear as two typical promising clean energy sources for ports. As a case study, the paper investigates the prospect of converting Alexandria Port in Egypt to be an eco-friendly port with the study of technical, logistic, and financial requirements. The results show that the fuel cell, followed by a combined system of wind turbines and fuel cells, is the best choice regarding electricity production unit cost by 0.101 and 0.107 $/kWh, respectively. Furthermore, using of fuel cells and offshore wind turbine as green power concept will achieving emissions reduction quantity of CO₂, NOx, and CO emissions by 80,441, 20.814, and 133.025 ton per year, respectively. Finally, the paper highlights the role that renewable energy can play when supplying Alexandria Port with green energy to lift the burden on the government in supporting the electricity, with a possibility of achieving a profit of 3.85% to 22.31% of the annual electricity cost compared with the international prices.

Keywords: fuel cells, green ports, IMO, national electric grid, offshore wind turbines, port emissions, renewable energy

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7640 Prevalence and Pathomorphological Study of Natural Coccidiosis in Japanese Quails (Coturnix coturnix japonica) in Iran

Authors: M. Khordadmehr, V. R. Ranjbar, R. Norouzi, M. Zeinoddin

Abstract:

Coccidiosis is recognized as a serious parasitic disease problem limiting quail industry recently. But the data on incidence, clinical signs, species of coccidia and pathological changes in Japanese quail are rare, especially in Iran in spite of the significant improvement of commercial quail breeding in this country in recent decades. Therefore, in the present paper was studied natural infection of quail coccidiosis in three commercial rearing farms with 80% morbidity and 3% mortality rate. For this purpose, fecal sample, oocyst examination, and morphological study were performed beside necropsy, histopathology, and PCR to confirm the diagnosis. In the affected birds, clinical signs included brown diarrhea, weakness, and pale face. In the fecal examination, three species of the genus Eimeria were identified including E. uzura, E. bateri, and E. tsunodai. At necropsy, the main gross lesions were edema, congestion and small blood spots in the small intestine. In histopathologic examination, endogenous stages of the parasites associated with hyperplasia of the intestinal glands, mild congestion, infiltration of mononuclear cells, and edema were observed in the intestine. The molecular study using BSEF and BSER specific primers confirmed the presence of the genus Eimeria in the affected birds. Interestingly, phylogenetic analysis showed relatively high bootstrap values in Japanese quail Eimeria with E. acervuline and E. maxima strains in the chicken. The present study is the first phylogenetic findings on Eimeria of quail which could be valuable for further research on Japanese quail coccidiosis.

Keywords: coccidiosis, Japanese Quail, pathomorphology, phylogenetic analysis

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7639 Studies on Radio Frequency Sputtered Copper Zinc Tin Sulphide Absorber Layers for Thin Film Solar Cells

Authors: G. Balaji, R. Balasundaraprabhu, S. Prasanna, M. D. Kannan, K. Sivakumaran, David Mcilroy

Abstract:

Copper Zin tin sulphide (Cu2ZnSnS4 or CZTS) is found to be better alternative to Copper Indium gallium diselenide as absorber layers in thin film based solar cells due to the utilisation of earth-abundant materials in the midst of lower toxicity. In the present study, Cu2ZnSnS4 thin films were prepared on soda lime glass using (CuS, ZnS, SnS) targets and were deposited by three different stacking orders, using RF Magnetron sputtering. The substrate temperature was fixed at 300 °C during the depositions. CZTS thin films were characterized using X-ray diffraction, X-ray photoelectron spectroscopy, Raman spectroscopy and UV-Vis-NIR spectroscopy. All the samples exhibited X-ray peaks pertaining to (112) kesterite phase of CZTS, along with the presence of a predominant wurtzite CZTS phase. X-ray photoelectron spectroscopy revealed the presence of all the elements in all the samples. The change in stacking order clearly shows that it affects the structural and phase properties of the films. Relative atomic concentrations of Zn, Cu, Sn and S, which are determined by high-resolution XPS core level spectra integrated peak areas revealed that the CZTS films exhibit inhomogeneity in both stoichiometry and elemental composition. Raman spectroscopy studies on the film showed the presence of CZTS phase. The energy band gap of the CZTS thin films was found to be in the range of 1.5 eV to 1.6 eV. The films were then annealed at 450 °C for 5 hrs and it was found that the predominant nature of the X-ray peaks has transformed from Wurtzite to Kesterite phase which is highly desirable for absorber layers in thin film solar cells. The optimized CZTS layer was used as an absorber layer in thin film solar cells. ZnS and CdS were used as buffer layers which in turn prepared by Hot wall epitaxy technique. Gallium doped Zinc oxide was used as a transparent conducting oxide. The solar cell structure Glass/Mo/CZTS/CdS or ZnS/GZO has been fabricated, and solar cell parameters were measured.

Keywords: earth-abundant, Kesterite, RF sputtering, thin film solar cells

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7638 Numerical Investigation of Phase Change Materials (PCM) Solidification in a Finned Rectangular Heat Exchanger

Authors: Mounir Baccar, Imen Jmal

Abstract:

Because of the rise in energy costs, thermal storage systems designed for the heating and cooling of buildings are becoming increasingly important. Energy storage can not only reduce the time or rate mismatch between energy supply and demand but also plays an important role in energy conservation. One of the most preferable storage techniques is the Latent Heat Thermal Energy Storage (LHTES) by Phase Change Materials (PCM) due to its important energy storage density and isothermal storage process. This paper presents a numerical study of the solidification of a PCM (paraffin RT27) in a rectangular thermal storage exchanger for air conditioning systems taking into account the presence of natural convection. Resolution of continuity, momentum and thermal energy equations are treated by the finite volume method. The main objective of this numerical approach is to study the effect of natural convection on the PCM solidification time and the impact of fins number on heat transfer enhancement. It also aims at investigating the temporal evolution of PCM solidification, as well as the longitudinal profiles of the HTF circling in the duct. The present research undertakes the study of two cases: the first one treats the solidification of PCM in a PCM-air heat exchanger without fins, while the second focuses on the solidification of PCM in a heat exchanger of the same type with the addition of fins (3 fins, 5 fins, and 9 fins). Without fins, the stratification of the PCM from colder to hotter during the heat transfer process has been noted. This behavior prevents the formation of thermo-convective cells in PCM area and then makes transferring almost conductive. In the presence of fins, energy extraction from PCM to airflow occurs at a faster rate, which contributes to the reduction of the discharging time and the increase of the outlet air temperature (HTF). However, for a great number of fins (9 fins), the enhancement of the solidification process is not significant because of the effect of confinement of PCM liquid spaces for the development of thermo-convective flow. Hence, it can be concluded that the effect of natural convection is not very significant for a high number of fins. In the optimum case, using 3 fins, the increasing temperature of the HTF exceeds approximately 10°C during the first 30 minutes. When solidification progresses from the surfaces of the PCM-container and propagates to the central liquid phase, an insulating layer will be created in the vicinity of the container surfaces and the fins, causing a low heat exchange rate between PCM and air. As the solid PCM layer gets thicker, a progressive regression of the field of movements is induced in the liquid phase, thus leading to the inhibition of heat extraction process. After about 2 hours, 68% of the PCM became solid, and heat transfer was almost dominated by conduction mechanism.

Keywords: heat transfer enhancement, front solidification, PCM, natural convection

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7637 Biosynthesis of a Nanoparticle-Antibody Phthalocyanine Photosensitizer for Use in Targeted Photodynamic Therapy of Cervical Cancer

Authors: Elvin P. Chizenga, Heidi Abrahamse

Abstract:

Cancer cell resistance to therapy is the main cause of treatment failures and the poor prognosis of cancer convalescence. The progression of cervical cancer to other parts of the genitourinary system and the reported recurrence rates are overwhelming. Current treatments, including surgery, chemo and radiation have been inefficient in eradicating the tumor cells. These treatments are also associated with poor prognosis and reduced quality of life, including fertility loss. This has inspired the need for the development of new treatment modalities to eradicate cervical cancer successfully. Photodynamic Therapy (PDT) is a modern treatment modality that induces cell death by photochemical interactions of light and a photosensitizer, which in the presence of molecular oxygen, yields a set of chemical reactions that generate Reactive Oxygen Species (ROS) and other free radical species causing cell damage. Enhancing PDT using modified drug delivery can increase the concentration of the photosensitizer in the tumor cells, and this has the potential to maximize its therapeutic efficacy. In cervical cancer, all infected cells constitutively express genes of the E6 and E7 HPV viral oncoproteins, resulting in high concentrations of E6 and E7 in the cytoplasm. This provides an opportunity for active targeting of cervical cancer cells using immune-mediated drug delivery to maximize therapeutic efficacy. The use of nanoparticles in PDT has also proven effective in enhancing therapeutic efficacy. Gold nanoparticles (AuNps) in particular, are explored for their use in biomedicine due to their biocompatibility, low toxicity, and enhancement of drug uptake by tumor cells. In this present study, a biomolecule comprising of AuNPs, anti-E6 monoclonal antibodies, and Aluminium Phthalocyanine photosensitizer was synthesized for use in targeted PDT of cervical cancer. The AuNp-Anti-E6-Sulfonated Aluminium Phthalocyanine mix (AlPcSmix) photosensitizing biomolecule was synthesized by coupling AuNps and anti-E6 monoclonal antibodies to the AlPcSmix via Polyethylene Glycol (PEG) chemical links. The final product was characterized using Transmission Electron Microscope (TEM), Zeta Potential, Uv-Vis Spectrophotometry, Fourier Transform Infrared Spectroscopy (FTIR), and X-ray diffraction (XRD), to confirm its chemical structure and functionality. To observe its therapeutic role in treating cervical cancer, cervical cancer cells, HeLa cells were seeded in 3.4 cm² diameter culture dishes at a concentration of 5x10⁵ cells/ml, in vitro. The cells were treated with varying concentrations of the photosensitizing biomolecule and irradiated using a 673.2 nm wavelength of laser light. Post irradiation cellular responses were performed to observe changes in morphology, viability, proliferation, cytotoxicity, and cell death pathways induced. Dose-Dependent response of the cells to treatment was demonstrated as significant morphologic changes, increased cytotoxicity, and decreased cell viability and proliferation This study presented a synthetic biomolecule for targeted PDT of cervical cancer. The study suggested that PDT using this AuNp- Anti-E6- AlPcSmix photosensitizing biomolecule is a very effective treatment method for the eradication of cervical cancer cells, in vitro. Further studies in vivo need to be conducted to support the use of this biomolecule in treating cervical cancer in clinical settings.

Keywords: anti-E6 monoclonal antibody, cervical cancer, gold nanoparticles, photodynamic therapy

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7636 Endothelin Cells and Its Molecular Biology and Microbiology

Authors: Chro Kawyan

Abstract:

Endothelin-1 (ET-1), the principal individual from the newfound mammalian endothelin group of organically dynamic peptides, was initially distinguished as a 21 buildup powerful vasoconstrictor peptide in vascular endothelial cells. However, it has since been demonstrated to have a wide range of pharmacological activities in tissues both inside and outside the cardiovascular system. Additionally, peptides that have a striking resemblance to ET-1 have been identified as the primary toxic component of snake venom. In addition, late examinations have proposed that warm blooded creatures, including people, produce three unmistakable individuals from this peptide family, ET-1, ET-2 and ET-J, which might have various profiles of organic action and may follow up on particular subtypes of endothelin receptor. Masashi Yanagisawa and Tomoh Masaki survey the ongoing status of the organic chemistry and sub-atomic science of endothelin.

Keywords: thelin, microbiology, molecular biology, cell

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7635 Molecular Identification of Camel Tick and Investigation of Its Natural Infection by Rickettsia and Borrelia in Saudi Arabia

Authors: Reem Alajmi, Hind Al Harbi, Tahany Ayaad, Zainab Al Musawi

Abstract:

Hard ticks Hyalomma spp. (family: Ixodidae) are obligate ectoparasite in their all life stages on some domestic animals mainly camels and cattle. Ticks may lead to many economic and public health problems because of their blood feeding behavior. Also, they act as vectors for many bacterial, viral and protozoan agents which may cause serious diseases such as tick-born encephalitis, Rocky-mountain spotted fever, Q-fever and Lyme disease which can affect human and/or animals. In the present study, molecular identification of ticks that attack camels in Riyadh region, Saudi Arabia based on the partial sequence of mitochondrial 16s rRNA gene was applied. Also, the present study aims to detect natural infections of collected camel ticks with Rickessia spp. and Borelia spp. using PCR/hybridization of Citrate synthase encoding gene present in bacterial cells. Hard ticks infesting camels were collected from different camels located in a farm in Riyadh region, Saudi Arabia. Results of the present study showed that the collected specimens belong to two species: Hyalomma dromedari represent 99% of the identified specimens and Hyalomma marginatum which account for 1 % of identified ticks. The molecular identification was made through blasting the obtained sequence of this study with sequences already present and identified in GeneBank. All obtained sequences of H. dromedarii specimens showed 97-100% identity with the same gene sequence of the same species (Accession # L34306.1) which was used as a reference. Meanwhile, no intraspecific variations of H. marginatum mesured because only one specimen was collected. Results also had shown that the intraspecific variability between individuals of H. dromedarii obtained in 92 % of samples ranging from 0.2- 6.6%, while the remaining 7 % of the total samples of H. dromedarii showed about 10.3 % individual differences. However, the interspecific variability between H. dromedarii and H. marginatum was approximately 18.3 %. On the other hand, by using the technique of PCR/hybridization, we could detect natural infection of camel ticks with Rickettsia spp. and Borrelia spp. Results revealed the natural presence of both bacteria in collected ticks. Rickettsial spp. infection present in 29% of collected ticks, while 35% of collected specimen were infected with Borrelia spp. The valuable results obtained from the present study are a new record for the molecular identification of camel ticks in Riyadh, Saudi Arabia and their natural infection with both Rickettsia spp. and Borrelia spp. These results may help scientists to provide a good and direct control strategy of ticks in order to protect one of the most important economic animals which are camels. Also results of this project spotlight on the disease that might be transmitted by ticks to put out a direct protective plan to prevent spreading of these dangerous agents. Further molecular studies are needed to confirm the results of the present study by using other mitochondrial and nuclear genes for tick identification.

Keywords: Camel ticks, Rickessia spp. , Borelia spp. , mitochondrial 16s rRNA gene

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7634 Effects of β-Glucan on the Release of Nitric Oxide by RAW264.7 Cells Stimulated with Escherichia coli Lipopolysaccharide

Authors: Eun Young Choi, So Hui Choe, Jin Yi Hyeon, Ji Young Jin, Bo Ram Keum, Jong Min Lim, Hyung Rae Cho, Kwang Keun Cho, In Soon Choi

Abstract:

This research analyzed the effect of β-glucan that is expected to alleviate the production of inflammatory mediator in macrophagocyte, which was processed by the lipopolysaccharide (LPS) of Escherichia, a pathogen related to allergy. The incubated layer was used for nitric oxide (NO) analysis. The DNA-binding activation of the small unit of NF-κB was measured using ELISA-based kit. In RAW264.7 cells that were vitalized by E.coli LPS, β-glucan inhibited both the combatant and rendering phases of inducible NO synthase (iNOS)-derived NO. β-glucan increased the expression of heme oxygenase-1 (HO-1) in the cell that was stimulated by E.coli LPS, and HO-1 activation was inhibited by SnPP. This shows that NO production induced by LPS is related to the inhibition effect of β-glucan. The phosphorylation of JNK and p38 induced by LPS were not influenced by β-glucan, and IκB-α decomposition was not influenced either. Instead, β-glucan remarkably inhibited the phosphorylation of STAT1 that was induced by E.coli LPS. Overall, β-glucan inhibited the production of NO in macrophagocyte that was vitalized by E.coli LPS through HO-1 induction and STAT1 pathways inhibition in this research. As the host inflammation reaction control by β-glucan weakens the progress of allergy, β-glucan can be used as an effective treatment method.

Keywords: β-glucan, lipopolysaccharide (LPS), nitric oxide (NO), RAW264.7 cells, STAT1

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7633 Evaluation of Collagen Synthesis in Macrophages/Fibroblasts Co-Culture Using Polylactic Acid Particles as Stimulants

Authors: Feng Ju Chuang, Yu Wen Wang, Tai Jung Hsieh, Shyh Ming Kuo

Abstract:

Polylactic acid is a synthetic polymer with good biocompatibility and degradability, is widely used in clinical applications. In this study, we utilized Polylactic acid particles as stimulants for macrophages and the collagen synthesis of co-cultured fibroblasts was evaluated. The results indicated that Polylactic acid particles were nontoxic to cells from 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. No obvious inflammation effect was observed (under the PLLA concentration of 1 mg/mL) after 24-h co-culture of Raw264.7 and NIH3T3 cells (from TNF-α assay). The addition of PLLA particles to the Raw264.7 and NIH3T3 co-cultures increased the synthesis of collagen, the highest collagen synthesis from the fibroblast was the 0.2 mg/mL (approximately 60% increased as compared with without addition Polylactic acid particles). Moreover, a co-axial atomization delivery device was used to percutaneously introduce Polylactic acid particles into the dermis layer and stimulating macrophages to secrete growth factors promoting fibroblasts to produce collagen. The preliminary results demonstrated the synthesis of collagen was increased mildly after the introduction of Polylactic acid particles for 28-d post implantation. The Polylactic acid particles could be successfully introduced into the dermis layer from H&E staining examination, however, the optimum concentration of Polylactic acid particles and the time-period for collagen synthesis still need to be evaluated.

Keywords: collagen synthesis, macrophage, NIH3T3 cells, polylactic acid particles

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7632 Optimization of Human Hair Concentration for a Natural Rubber Based Composite

Authors: Richu J. Babu, Sony Mathew, Sharon Rony Jacob, Soney C. George, Jibin C. Jacob

Abstract:

Human hair is a non-biodegradable waste available in plenty throughout the world but is rarely explored for applications in engineering fields. Tensile strength of human hair ranges from 170 to 220 MPa. This property of human hair can be made use in the field of making bio-composites[1]. The composite is prepared by commixing the human hair and natural rubber in a two roll mill along with additives followed by vulcanization. Here the concentration of the human hair is varied by fine-tuning the fiber length as 20 mm and sundry tests like tensile, abrasion, tear and hardness were conducted. While incrementing the fiber length up to a certain range the mechanical properties shows superior amendments.

Keywords: human hair, natural rubber, composite, vulcanization, fiber loading

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7631 Binding Mechanism of Synthesized 5β-Dihydrocortisol and 5β-Dihydrocortisol Acetate with Human Serum Albumin to Understand Their Role in Breast Cancer

Authors: Monika Kallubai, Shreya Dubey, Rajagopal Subramanyam

Abstract:

Our study is all about the biological interactions of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules with carrier protein Human Serum Albumin (HSA). The cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 µM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. The further experiment proved that Dhc and DhcA induced 35.6% and 37.7% early apoptotic cells and 2.5%, 2.9% late apoptotic cells respectively. Morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7±0.03×104 M-1 and 3.9±0.05×104 M-1, and their binding free energies were found to be -6.4 and -6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4±0.05×104 M-1 replaced Dhc, and phenylbutazone 1.5±0.05×104 M-1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular sizes of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported the greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for the further development of steroid derivatives with improved pharmacological significance as novel anti-cancer drugs.

Keywords: apoptosis, dihydrocortisol, fluorescence quenching, protein conformations

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7630 Isolation and Culture of Keratinocytes and Fibroblasts to Develop Artificial Skin Equivalent in Cats

Authors: Lavrentiadou S. N., Angelou V., Chatzimisios K., Papazoglou L.

Abstract:

The aim of this study was the isolation and culture of keratinocytes and fibroblasts from feline skin to ultimately create an artificial engineered skin (including dermis and epidermis) useful for the effective treatment of large cutaneous deficits in cats. Epidermal keratinocytes and dermal fibroblasts were freshly isolated from skin biopsies using an 8 mm biopsy punch obtained from 8 healthy cats that had undergone ovariohysterectomy. The owner’s consent was obtained. All cats had a complete blood count and a serum biochemical analysis and were screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) preoperatively. The samples were cut into small pieces and incubated with collagenase (2 mg/ml) for 5-6 hours. Following digestion, cutaneous cells were filtered through a 100 μm cell strainer, washed with DMEM, and grown in DMEM supplemented with 10% FBS. The undigested epidermis was washed with DMEM and incubated with 0.05% Trypsin/0.02% EDTA (TE) solution. Keratinocytes recovered in the TE solution were filtered through a 100 μm and a 40 μm cell strainer and, following washing, were grown on a collagen type I matrix in DMEM: F12 (3:1) medium supplemented with 10% FΒS, 1 μm hydrocortisone, 1 μm isoproterenol and 0.1 μm insulin. Both fibroblasts and keratinocytes were grown in a humidified atmosphere with 5% CO2 at 37oC. The medium was changed twice a week and cells were cultured up to passage 4. Cells were grown to 70-85% confluency, at which point they were trypsinized and subcultured in a 1:4 dilution. The majority of the cells in each passage were transferred to a freezing medium and stored at -80oC. Fibroblasts were frozen in DMEM supplemented with 30% FBS and 10% DMSO, whereas keratinocytes were frozen in a complete keratinocyte growth medium supplemented with 10% DMSO. Both cell types were thawed and successfully grown as described above. Therefore, we can create a bank of fibroblasts and keratinocytes, from which we can recover cells for further culture and use for the generation of skin equivalent in vitro. In conclusion, cutaneous cell isolation and cell culture and expansion were successfully developed. To the authors’ best knowledge, this is the first study reporting isolation and culture of keratinocytes and fibroblasts from feline skin. However, these are preliminary results and thus, the development of autologous-engineered feline skin is still in process.

Keywords: cat, fibroblasts, keratinocytes, skin equivalent, wound

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7629 Quantification and Evaluation of Tumors Heterogeneity Utilizing Multimodality Imaging

Authors: Ramin Ghasemi Shayan, Morteza Janebifam

Abstract:

Tumors are regularly inhomogeneous. Provincial varieties in death, metabolic action, multiplication and body part are watched. There’s expanding proof that strong tumors may contain subpopulations of cells with various genotypes and phenotypes. These unmistakable populaces of malignancy cells can connect during a serious way and may contrast in affectability to medications. Most tumors show organic heterogeneity1–3 remembering heterogeneity for genomic subtypes, varieties inside the statement of development variables and genius, and hostile to angiogenic factors4–9 and varieties inside the tumoural microenvironment. These can present as contrasts between tumors in a few people. for instance, O6-methylguanine-DNA methyltransferase, a DNA fix compound, is hushed by methylation of the quality advertiser in half of glioblastoma (GBM), adding to chemosensitivity, and improved endurance. From the outset, there includes been specific enthusiasm inside the usage of dissemination weighted imaging (DWI) and dynamic complexity upgraded MRI (DCE-MRI). DWI sharpens MRI to water dispersion inside the extravascular extracellular space (EES) and is wiped out with the size and setup of the cell populace. Additionally, DCE-MRI utilizes dynamic obtaining of pictures during and after the infusion of intravenous complexity operator. Signal changes are additionally changed to outright grouping of differentiation permitting examination utilizing pharmacokinetic models. PET scan modality gives one of a kind natural particularity, permitting dynamic or static imaging of organic atoms marked with positron emanating isotopes (for example, 15O, 18F, 11C). The strategy is explained to a colossal radiation portion, which points of confinement rehashed estimations, particularly when utilized together with PC tomography (CT). At long last, it's of incredible enthusiasm to quantify territorial hemoglobin state, which could be joined with DCE-CT vascular physiology estimation to create significant experiences for understanding tumor hypoxia.

Keywords: heterogeneity, computerized tomography scan, magnetic resonance imaging, PET

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